Sample analyzer and computer program product

ABSTRACT

A sample analyzer prepares a measurement sample from a blood sample or a body fluid sample which differs from the blood sample; measures the prepared measurement sample; obtains characteristic information representing characteristics of the components in the measurement sample; sets either a blood measurement mode for measuring the blood sample, or a body fluid measurement mode for measuring the body fluid sample as an operating mode; and measures the measurement sample prepared from the blood sample by executing operations in the blood measurement mode when the blood measurement mode has been set, and measuring the measurement sample prepared from the body fluid sample by executing operations in the body fluid measurement mode that differs from the operations in the blood measurement mode when the body fluid measurement mode has been set, is disclosed. A computer program product is also disclosed.

This application is a Continuation of U.S. application Ser. No.13/891,667 filed May 10, 2013, which is a Continuation of U.S.application Ser. No. 12/023,830 filed Jan. 31, 2008, now U.S. Pat. No.8,440,140, claiming priority to Japanese Application No. 2007-022523filed on Feb. 1, 2007 and to Japanese Application No. 2007-095226 filedon Mar. 30, 2007, all of which are incorporated herein by reference intheir entireties.

FIELD OF THE INVENTION

The present invention relates to a sample analyzer and a computerprogram product capable of measuring not only blood, but also bodyfluids other than blood such as cerebrospinal fluid (spinal fluid),fluid of the thoracic cavity (pleural fluid), abdominal fluid and thelike.

BACKGROUND

In the field of clinical examinations, blood is routinely collected froma body and used as a sample which is measured by a sample analyzer toaid diagnosis and monitor treatment. Furthermore, body fluids other thanblood are also often used as samples which are measured by a sampleanalyzer. The body fluids are usually transparent and contain very fewcells, however, cells such as bacteria, abnormal cells, and hemorrhage(blood cells) and the like may be found in cases of disease, tumors ofrelated organs, and injury.

When cerebrospinal fluid, which is one type of body fluid, is measured,for example, it is possible to make the following estimations from themeasurement results.

-   -   Increase of red blood cells: subarachnoidal hemorrhage    -   Increase of neutrophils: meningitis    -   Increase of eosinophils: infectious disease (parasites and        fungus)    -   Increase of monocytes: tuberculous meningitis, viral meningitis    -   Other cells: advanced meningeal tumor

Japanese Laid-Open Patent Publication No. 2003-344393 discloses a bloodcell analyzer which is capable of measuring cells in a body fluid. InJapanese Laid-Open Patent Publication No. 2003-344393, an operatorprepares a measurement sample prior to performing the measurements bymixing a fluid sample and reagent (aldehyde, surface active agent, andcyclodextrin) in order to stably store the body fluid for a long period,and this measurement sample is later subjected to fluid analysis by thesample analyzer.

In the art of Japanese Laid-Open Patent Publication No. 2003-344393,however, the measurement sample is not prepared by the sample analyzerwhen the body fluid is measured, rather the measurement sample must beprepared by the operator of the analyzer. Furthermore, the sampleanalyzer disclosed in Japanese Laid-Open Patent Publication No.2003-344393 does not disclose measurement operations suited to the fluidwhen measuring a body fluid.

SUMMARY OF THE INVENTION

The scope of the present invention is defined solely by the appendedclaims, and is not affected to any degree by the statements within thissummary.

A first aspect of the present invention is a sample analyzer comprising:a measuring part for preparing a measurement sample from a blood sampleor a body fluid sample that differs from the blood sample, measuring theprepared measurement sample, and obtaining characteristic informationrepresenting characteristics of components within the measurementsample; a mode setting means for setting either a blood measurement modefor measuring the blood sample, or a body fluid measurement mode formeasuring the body fluid sample as an operating mode; a first controlmeans for controlling the measuring part so as to execute operations inthe blood measurement mode when the blood measurement mode has been setby the mode setting means; and a second control means for controllingthe measuring part so as to execute operations in the body fluidmeasurement mode which differs from the operations in the bloodmeasurement mode when the body fluid measurement mode has been set bythe mode setting means.

A second aspect of the present invention is a sample analyzercomprising: a measuring part for preparing a measurement sample from ablood sample or a body fluid sample that differs from the blood sample,measuring the prepared measurement sample, and obtaining characteristicinformation representing characteristics of components within themeasurement sample; a mode setting means for setting either a bloodmeasurement mode for measuring the blood sample, or a body fluidmeasurement mode for measuring the body fluid sample as an operatingmode; a first analyzing means for executing a first analysis processbased on the characteristic information obtained by measuring themeasurement sample prepared by the measuring part from the blood samplewhen the blood measurement mode has been set by the mode setting means;and a second analyzing means for executing a second analysis processwhich differs from the first analysis process based on thecharacteristic information obtained by measuring the measurement sampleprepared by the measuring part from the body fluid sample when the bodyfluid measurement mode has been set by the mode setting means.

A third aspect of the present invention is a sample analyzer comprising:a measuring part for preparing a measurement sample from a blood sampleor a body fluid sample that differs from the blood sample, measuring theprepared measurement sample, and obtaining characteristic informationrepresenting characteristics of components within the measurementsample; a mode switching means for switching an operating mode from ablood measurement mode for measuring the blood sample to a body fluidmeasurement mode for measuring the body fluid sample; and a blankmeasurement controlling means for controlling the measuring part so asto measure a blank sample that contains neither the blood sample nor thebody fluid sample when the mode switching means has switched theoperating mode from the blood measurement mode to the body fluidmeasurement mode.

A fourth aspect of the present invention is a computer program product,comprising: a computer readable medium; and instructions, on thecomputer readable medium, adapted to enable a general purpose computerto perform operations, comprising: a step of preparing a measurementsample from a blood sample or a body fluid sample which differs from theblood sample; a step of measuring the prepared measurement sample; astep of obtaining characteristic information representingcharacteristics of the components in the measurement sample; a step ofsetting either a blood measurement mode for measuring the blood sample,or a body fluid measurement mode for measuring the body fluid sample asan operating mode; and a step of measuring the measurement sampleprepared from the blood sample by executing operations in the bloodmeasurement mode when the blood measurement mode has been set, andmeasuring the measurement sample prepared from the body fluid sample byexecuting operations in the body fluid measurement mode that differsfrom the operations in the blood measurement mode when the body fluidmeasurement mode has been set.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an exterior view of a blood cell analyzer of a firstembodiment of the present invention;

FIG. 2 is a block diagram of the measuring unit of the analyzer;

FIG. 3 is a block diagram of the fluid supplying unit;

FIG. 4 shows the optical system of the white blood cell detection unit;

FIG. 5 shows the RBC/PLT detection unit;

FIG. 6 shows the HGB detection unit;

FIG. 7 is a flow chart of the sample measuring process;

FIG. 8 shows the display screen for setting the measurement mode;

FIG. 9 is a flow chart showing the pre sequence process;

FIG. 10 is a schematic view of a scattergram derived from measurementsof a DIFF measurement sample prepared from body fluid;

FIG. 11 compares measurement results by the blood cell analyzer of theembodiment and measurement results by a reference method;

FIG. 12 is a schematic view of a scattergram derived from measurementsof a DIFF measurement sample prepared from blood;

FIG. 13 is a display screen showing the measurement results in the bloodmeasurement mode;

FIG. 14 is a display screen showing the measurement results in the bodyfluid measurement mode;

FIG. 15 is a display screen showing the measurement results in the bodyfluid measurement mode;

FIG. 16 is a display screen showing the measurement results in the bodyfluid measurement mode; and

FIG. 17 is a confirmation screen at the start of the blank check whichis displayed in the body fluid measurement mode.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The preferred embodiments of the present invention will be describedhereinafter with reference to the drawings.

FIG. 1 shows a sample analyzer 1. The sample analyzer 1 is configured asan automatic multi-item blood cell analyzer which performs bloodanalysis by measuring blood samples held in sample containers (bloodcollection tubes), obtaining characteristics information representingthe characteristics of the blood cells contained in the sample, andanalyzing the characteristic information. The sample analyzer 1 is alsocapable of analyzing body fluids. In the blood cell analyzer of thepresent embodiment, the body fluids used as analysis objects include,fluid within the body cavity other than blood. Specifically,cerebrospinal fluid (spinal fluid, CSF: fluid filling the ventricle orsublemmal cavity), fluid of the thoracic cavity (pleural fluid, PE:fluid collected in pleural cavity), abdominal fluid (fluid collected inthe abdominal cavity), fluid of the cardiac sac (fluid collected in thecardiac sac), synovial fluid (fluid present in joints, synovial sac,peritenon) and the like. Among types of body fluid which can be analyzedare dialysate of peritoneal dialysis (CAPD), intraperitoneal rinse andthe like. Cells are usually not observed in these body fluids, however,the fluids may contain blood cells, abnormal cells, and cells such asbacteria in the case of disease, tumor of related organs, or injury. Forexample, it is possible to clinically estimate the following frommeasurement results in the case of cerebrospinal fluid. For example,sub-arachnoidal hemorrhage is indicted when there is an increase of redblood cells, meningitis is indicated when there is an increase ofneutrophils, infectious disease (parasitic and fungal) is indicated whenthere is an increase of eosinophils, tuberculous meningitis and viralmeningitis are indicated when there is an increase of monocytes, andadvanced meningeal tumor is indicated when there is an increase of othercells. ed In the case of abdominal and thoracic fluids, cancers may beindicated when analysis of finds nucleated cells other than blood cells,that is, the fluid contains nucleated cells of mesothelial cells,macrophages, tumor cells and the like.

The sample analyzer 1 is provided with a measuring unit 2 which has thefunction of measuring blood and body fluid samples, and a dataprocessing unit 3 which obtains analysis results by processing themeasurement results output from the measurement unit 2. The dataprocessing unit 3 is provided with a control unit 301, a display unit302, and an input unit 303. Although the measuring unit 2 and dataprocessing unit 3 are separate devices in FIG. 1, the both may also beintegrated in a single apparatus.

FIG. 2 is a block diagram of the measuring unit 2 of the analyzer 1. Asshown in FIG. 2, the measuring unit 2 is provided with a blood celldetecting unit 4, an analog processing unit 5 which processes the output(analog signals) of the detecting unit 4, microcomputer unit 6, displayand operating unit 7, and a device 8 for measuring blood and bodyfluids. The device 8 includes a fluid supplying unit 81 which isdescribed below.

FIG. 3 is a block diagram showing the structure of the fluid supplyingunit 81. As shown in FIG. 3, the fluid supplying unit 81 is providedwith a sample aspiration nozzle 18, a plurality of reagent containers, asampling valve 12, and reactions chambers 13 through 17. The sampleaspiration nozzle 18 aspirates sample from a sample container, anddelivers the sample to the sampling valve 12. The sampling valve 12divides the delivered sample into several aliquots of predeterminedvolume. The number of divisions differs depending on the mode ofmeasurement (discrete mode); in the CBC mode the sample is divided intothree aliquots to measure the number of red blood cells, the number ofwhite blood cells, the number of platelets, and the hemoglobinconcentration. In addition to the CBC measurement items, the sample isdivided into four aliquots in the CBC-DIFF mode so as to also classifyfive types of white blood cells. Furthermore, In addition to themeasurement items of the CBC+DIFF mode, the sample is divided into fivealiquots in the CBC+DIFF+RET mode so as to also measure reticulocytes.

Similarly, in addition to the measurement items of the CBC+DIFF mode,the sample is divided into five aliquots in the CBC+DIFF+NRBC mode so asto also measure nucleated red blood cells. In addition to themeasurement items of the CBC+DIFF+RET mode, the sample is divided intosix aliquots in the CBC+DIFF+RET+NRBC mode so as to also measurenucleated red blood cells. The above mentioned measurement modes areblood measuring modes which measure whole blood. Finally, the sample isdivided into two aliquots in the body fluid measuring mode for measuringbody fluid.

Reagent (dilution solution) is introduced from a reagent container tothe sampling valve, and the aliquots of the divided sample are deliveredtogether with the reagent to the reaction chambers 13 through 17 and anHGB detection unit 43, which is described later. a predetermined amountof sample (aliquot) and a predetermined amount of reagent and apredetermined amount of stain collected by the sampling valve 12 aresupplied to the reaction chamber 13 by a dosage pump which is not shownin the drawing, the sample and reagent are mixed to prepare ameasurement sample for four classifications of white blood cells (DIFF).

The reagent “stomatolyzer 4DL” made by Sysmex Corporation may be used asthe dilution solution. This reagent contains surface active agent andinduces hemolysis of red blood cells. The reagent “stomatolyzer 4DS”made by Sysmex Corporation may be used as the stain. This stain containsethylene glycol, low molecular alcohol, and polymethene colorant; a 50×dilute sample is ultimately prepared by staining the blood cellcomponent after hemolysis by the dilution agent.

When the body fluid measurement mode has been selected, a measurementsample for the classification of white blood cells is prepared from afluid sample under the conditions of the amount of the sample andreagent used for the four classifications of white blood cells areidentical, the reagents are identical, and the amounts of the reagentare identical. In the white blood cell classification of the body fluidmeasurement mode, the white blood cells are classified, not in fourtypes, but two types, as shall be described later.

A predetermined amount of sample collected by the sampling valve 12, apredetermined amount of hemolytic dilution agent, and a predeterminedamount of stain solution are supplied to the reaction chamber 14 by adosage pump which is not shown in the drawing, the sample and reagentsare then mixed to prepare a measurement sample for measuring nucleatedred blood cells (NRBC).

A predetermined amount of sample collected by the sampling valve 12, apredetermined amount of dilution agent, and a predetermined amount ofstain solution are supplied to the reaction chamber 15 by a dosage pumpwhich is not shown in the drawing, the sample and reagents are thenmixed to prepare a measurement sample for measuring reticulocytes (RET).

A predetermined amount of sample collected by the sampling valve 12, anda predetermined amount of hemolytic dilution agent are supplied to thereaction chamber 16 by a dosage pump which is not shown in the drawing,the sample and reagents are then mixed to prepare a measurement samplefor measuring white blood cells and basophils (WBC/BASO).

A predetermined amount of sample collected by the sampling valve 12, anda predetermined amount of dilution solution are supplied to the reactionchamber 17 by a dosage pump which is not shown in the drawing, thesample and reagents are then mixed to prepare a measurement sample formeasuring red blood cells and platelets (RBC/PLT).

A predetermined amount of sample collected by the sampling valve 12, anda predetermined amount of hemolytic dilution agent are supplied to theHGB detection unit 43 which is described later.

The detection device 4 is provided with a white blood cell detectionunit 41 for detecting white blood cells. The white blood cell detectionunit 41 is also used to detect nucleated red blood cells andreticulocytes. In addition to the white blood cell detection unit, thedetection device 4 is also provided with an RBC/PLT detection unit 42for measuring the number of red blood cells and the number of platelets,and an HGB detection unit 43 for measuring the amount of pigment in theblood.

The white blood cell detection unit 41 is configured as an opticaldetection unit, specifically, a detection unit which uses a flowcytometric method. Cytometry measures the optical properties andphysical properties of cells and other biological particles, and flowcytometry measures these particles as they pass by in a narrow flow.FIG. 4 shows the optical system of the white blood cell detection unit41. In the same drawing, the beam emitted from a laser diode 401irradiates, via a collimator lens 402, the blood cells passing throughthe interior of a sheath flow cell 403. The intensity of the frontscattered light, the intensity of the side scattered light, and theintensity of the side fluorescent light from the blood cells within thesheath flow cell irradiated by the light are detected by the white bloodcell detection unit 41.

The scattered light is a phenomenon due to the change in the directionof travel of the light caused by particles such as blood cells and thelike which are present as obstructions in the direction of travel of thelight. Information on the characteristics of the particles related tothe size and composition of the particles can be obtained by detectingthis scattered light. The front scattered light emerges from theparticles in approximately the same direction as the direction of travelof the irradiating light. Characteristic information related to the sizeof the particle (blood cell) can be obtained from the front scatteredlight. The side scattered light emerges from the particle in anapproximate perpendicular direction relative to the direction of travelof the irradiating light. Characteristic information related to theinterior of the particle can be obtained from the side scattered light.When a particle is irradiated by laser light, the side scattered lightintensity is dependent on the complexity (that is, nucleus shape, size,density, and granularity) of the interior of the cell. therefore, theblood cells can be classified (discriminated) and the number of cellscan be counted by using the characteristics of the side scattered lightintensity. Although the front scattered light and side scattered lightare described as the scattered light used in the present embodiment, thepresent invention is not limited to this configuration inasmuch asscattered light of any angle may also be used relative to the opticalaxis of the light emitted from a light source that passes through thesheath flow cell insofar as scattered light signals are obtained whichrepresent the characteristics of the particles necessary for analysis.

When fluorescent material such as a stained blood cell is irradiated bylight, light is given off by the particle at a wavelength which islonger than the wavelength of the irradiating light. The intensity ofthe fluorescent light is increased by the stain, and characteristicsinformation can be obtained relating to the degree of staining of theblood cell by measuring the fluorescent light intensity. Theclassification and other measurements of the white blood cells can thenbe performed by the difference in the (side) fluorescent lightintensity.

As shown in FIG. 4, the front scattered light from the blood cell (whiteblood cells and nucleated red blood cells) which pass through the sheathflow cell 403 is received by a photodiode (front scattered lightreceiving unit) 406 through a collective lens 404 and pinhole 405. Theside scattered light is received by a photo multiplexer (side scatteredlight receiving unit) 411 through a collective lens 407, dichroic mirror408, optical filter 409, and pinhole 410. The side fluorescent light isreceived by a photo multiplexer (side fluorescent light receiving unit)412 through the collective lens 407 and dichroic mirror 408. Thephotoreception signals output from the light receiving units 406, 411,and 412 are subjected to analog processing such as amplification andwaveform processing and the like by an analog processing unit 5 which isconfigured by amps 51, 52, 53 and the like, and the analog-processedphotoreception signals are provided to the microcomputer 6.

The configuration of the RBC/PLT detection unit 42 is described below.FIG. 5 is a schematic view briefly showing the structure of the RBC/PLTdetection unit 42. The RBC/PLT detection unit 42 is capable of measuringthe numbers of red blood cells and platelets by a sheath flow-DCdetection method. The RBC/PLT detection unit 42 has a sheath flow cell42 a as shown in FIG. 5. The sheath flow cell 42 a is provided with asample nozzle 42 b which is open toward the top so that sample can besupplied from the reaction chamber 17 to the sample nozzle 42 b. Thesheath flow cell 42 a has a tapered chamber 42 c which narrows towardthe top, and the sample nozzle 42 b is disposed in the center partwithin the chamber 42 c. An aperture 42 d is provided at the top end ofthe chamber 42 c, and this aperture 42 d is aligned with the centerposition of the sample nozzle 42 b. Measurement sample supplied from thesample supplying unit is sent upward from the tip of the sample nozzle42 b, and front sheath fluid is simultaneously supplied to the chamber42 c and flows upward toward the aperture 42 d. The flow of themeasurement sample, which is encapsulated in the front sheath fluid, isnarrowly constricted by the tapered chamber 42 c and the blood cellswithin the measurement sample pass one by one through the aperture 42 d.Electrodes are provided at the aperture 42 d, and a direct current issupplied between these electrodes. The change in the resistance of thedirect current is detected at the aperture 42 d when the measurementsample flows through the aperture 42 d, and the electrical signal of thechange in resistance is output to the controller 25. Since theresistance of the direct current increases when blood cells pass throughthe aperture 42 d, the electrical signals reflect information of thepassage of the blood cells through the aperture 42 d so that the numbersof red blood cells and platelets can be counted by subjecting theseelectrical signals to signal processing.

A recovery tube 42 e, which extends vertically, is provided above theaperture 42 d. The recovery tube 42 e is disposed within a chamber 42 fwhich is connected to the chamber 42 c through the aperture 42 d. Theinner wall of the chamber 42 f is separated from the bottom end of therecovery tube 42 e. The chamber 42 f is configured to supply a backsheath, and this back sheath flows downward through the chamber 42 f ina region outside the recovery tube 42 e. The back sheath which flowsoutside the recovery tube 42 e arrives at the bottom part of the chamber42 f, and thereafter flows between the inner wall of the chamber 42 fand the bottom end of the recovery tube 42 e so as to flow into theinterior of the recovery tube 42 e. The blood cells which has passedthrough the aperture 42 d are therefore prevented from refluxing, thuspreventing erroneous detection of the blood cells.

The configuration of the HGB detection unit 43 is described below. TheHGB detection unit 43 is capable of measuring the amount of hemoglobin(HGB) by an SLS hemoglobin method. FIG. 6 is a perspective view of thestructure of the HGB detection unit 43. The HGB detection unit 43 has acell 43 a for accommodating a diluted sample, a light-emitting diode 43b for emitting light toward the cell 43 a, and a photoreceptor element43 c for receiving the transmission light that has passed through thecell 43 a. A fixed amount of blood is diluted with dilution fluid and apredetermined hemolytic agent at a predetermined dilution ratio by thesampling valve 12 to prepare a dilute sample. The hemolytic agent hasproperties which transform the hemoglobin in the blood toSLS-hemoglobin. The dilute sample is supplied to the cell 43 a andaccommodated therein. In this condition, the light-emitting diode 43 bemits light that passes through the cell 43 a and is received by thephotoreceptor element 43 c which is disposed opposite the light-emittingdiode 43 b with the cell 43 a interposed therebetween. Since thelight-emitting diode 43 b emits light having a wavelength that is highlyabsorbed by the SLS-hemoglobin, and the cell 43 a is configured ofplastic material which has a high light transmittancy, the photoreceptorelement 43 c only receives the transmission light absorbed by the dilutesample of the light emitted from the light-emitting diode 43 b. Thephotoreceptor element 43 c outputs electrical signals which correspondto the amount of received light (optical density) to the microcomputer6, and the microcomputer 6 compares the optical density with the opticaldensity of the dilution solution which was measured previously, thencalculates the hemoglobin value.

The microcomputer 6 is provided with an A/D converter 61 for convertingthe analog signals received from the analog processing unit 5 to digitalsignals. The output of the A/D converter 61 is sent to a calculationunit 62 of the microcomputer 6, and calculations are performed forpredetermined processing of the photoreception signals in thecalculation unit 62. The calculation unit 62 prepares distribution data(two-dimensional scattergrams (unclassified) and unidimensionalhistograms) based on the output of the detection device 4.

The microcomputer 6 is provided with a controller 63 configured by amemory for the control processor and the operation of the controlprocessor, and a data analyzing unit 64 configured by a memory for theanalysis processor and the operation of the analysis processor. Thecontroller 63 controls the device 8 configured by a sampler (not shownin the drawing) for automatically supplying blood collection tubes, anda fluid system and the like for preparing and measuring samples, as wellas performing other controls. The data analyzing unit 64 executesanalysis processing such as clustering and the like on the distributiondata. The analysis results are sent to an external data processingdevice 3 through an interface 65, and the data processing device 3processes the data for screen display, storage and the like.

The microcomputer 6 is further provided with an interface 66 which isinterposed between the microcomputer 6 and the display and operatingunit 7, and an interface 67 which is interposed between themicrocomputer 6 and the device 8. The calculation unit 62, controller63, and interfaces 66 and 67 are connected through a bus 68, and thecontroller 63 and the data analyzing unit 64 are connected through a bus69. The display and operating unit 7 includes a start switch by whichthe operator specifies to start a measurement, and a touch panel typeliquid crystal display for displaying various types of setting valuesand analysis results, and receiving input from the operator.

The operation of the sample analyzer 1 of the present embodiment isdescribed below. FIG. 7 is a flow chart showing the flow of theoperation of the sample analyzer of the present embodiment. The sampleanalyzer 1 starts when a user turns on the power source of the sampleanalyzer 1 (step S1). The sample analyzer 1 first executes a self checkduring startup (step S2). In the self check, the microcomputer 6 testsand checks the operation of all operating device of the sample analyzer1, and performs a blank check operation which measures a blank samplethat does not contain a real sample. Next, the microcomputer 6 sets aninitial measurement mode (step S3). The CBC+DIFF mode is the initialsetting. Specifically, in the process of step S3, parameters (operatingconditions) for performing blood measurements are set, for example,which reaction chamber to use and the set time for the measurement. Theblood measurement mode is thus set as the initial operating mode in thesample analyzer 1 of the present embodiment. The sample analyzer 1therefore remains in a standby state waiting to receive a measurementstart instruction. The microcomputer 6 displays a screen on the liquidcrystal display which alerts the operator to the standby state (stepS4).

In the standby state, the operator can change the measurement mode byoperating the display and operation unit 7. FIG. 8 is a schematic viewof an input screen for setting the measurement mode. This screen isprovided with discrete display regions including the sample number 120,type of sample uptake mode 121, type of discrete test (measurement mode)122, and type of sample 123. The three sample uptake modes include amanual mode for aspirating a sample after the operator has manuallyinserted a sample container in the sample aspiration nozzle 18, acapillary mode for aspirating a measurement sample via the sampleaspiration nozzle 18 after the operator has previously prepared themeasurement sample by mixing a sample and reagent, and a closed mode forsupplying a sample by automatically transporting a sample containerusing a conveyer device. The types of samples include NORMAL, which arenormal blood samples; HPC, which are hematopoietic progenitor cellsamples; and BODY FLUID, which are other fluids of the body. Theoperator can specify the sample take-up mode, measurement mode, and typeof sample. When the blood measurement mode has been specified, theNORMAL sample type is specified, and an optional sample take-up mode andmeasurement mode are specified. When specifying the BODY FLUIDmeasurement mode, the operator specifies MANUAL mode as the take-upmode, [CBC+DIFF], [CBC+DIFF+RET], [CBC+DIFF+NRBC], or [CBC+DIFFNRBC+RET]as the DISCRETE test, and [BODY FLUID] as the type of sample. In stepS4, the operator specifies the desired mode. The operator presses thestart switch to start the measurement when blood measurement isperformed without changing the initially set measurement mode (step S5:N). The microcomputer 6 receives the instruction to start themeasurement (step S6), and the blood sample is aspirated by the sampleaspiration nozzle (step S7).

After the blood sample has been aspirated, the sample is introduced tothe previously mentioned sampling valve 18, and the necessary samplepreparation is performed for the measurement according to the typediscrete test of the measurement mode (step S14). The measurementoperation is then executed for this measurement sample (step S16).When[7] is set as the type of discrete test, for example, HGB, WBC/BASO,DIFF, RET, NRBC, and RBC/PLT measurement samples are prepared.Thereafter, the WBC/BASO, DIFF, RET, and NRBC measurement samples aremeasured by the white blood cell detection unit 41, the RBC/PLTmeasurement sample is measured by the RBC/PLT detection unit 42, and theHGB measurement sample is measured by the HGB detection unit 43. At thistime, the WBC/BASO, DIFF, RET, and NRBC measurement samples areintroduced to the white blood cell detection unit 41 in the order NRBC,WBC/BASO, DIFF, RET and sequentially measured since only a single whiteblood cell detection unit 41 is provided. In this measurement operation,the calculation unit 62 creates particle distribution maps (scattergram,histogram). The scattergram created from the optical informationobtained by the DIFF measurement is described below. The calculationunit 62 generates a two-dimensional scattergram (particle distributionmap) using, as characteristic parameters, the side scattered light andside fluorescent light among the photoreception signals output from thewhite blood cell detection unit 41 in the DIFF measurement. Thisscattergram (referred to as “DIFF scattergram” hereinafter) plots theside scattered light intensity on the X axis and the side fluorescentlight on the Y axis; red blood cell ghost clusters, lymphocyte clusters,monocyte clusters, neutrophil+basophil clusters, and eosinophil clustersnormally appear. These clusters are recognized by processing performedon the DIFF scattergram by the data analyzing unit 64.

Analysis processing is then performed based on the particle distributionmaps obtained by the measurement (step S18). In the analysis processing,the data analyzing unit 64 of the microcomputer 6 classifies the fourwhite blood cell clusters (lymphocyte cluster, monocyte cluster,neutrophil+basophil cluster, and eosinophil cluster), and the red bloodcell ghost cluster as shown in FIG. 12 from the DIFF scattergramprepared by the calculation unit 62 when the DIFF measurement sampleswere measured by the white blood cell detection unit 41. In the analysisprocess of the present embodiment, each particle plotted on thescattergram and the degree of attribution of particles to each clusterat a distance from the center of gravity of each cluster is obtained.Then, each particle is attributed to a cluster according to the degreeof attribution. The particle classification method is disclosed indetail in U.S. Pat. No. 5,555,196. The basophil cluster, and white bloodcell clusters other than basophils, and the red blood cell ghost clusterare classified on the scattergram obtained by the WBC/BASO measurement.White blood cells are classified in five groups based on the results ofthe four classifications and numbers of white blood cells (refer to FIG.12) by the analysis processing of the DIFF scattergram, and the resultsof the two classification and numbers of white blood cells by theanalysis processing of the WBC/BASO scattergram. Specifically, the dataanalysis unit 64 subtracts the basophil cell count obtained by theanalyzing the WBC/BASO scattergram from the neutrophil+basophil cellcount obtained by analyzing the DIFF scattergram, to obtain theneutrophil cell count and the basophil cell count. Thus, fiveclassifications of white blood cells are obtained as well as the numberof blood cells in each classification. In addition, the trough isdetected in the curve in the unidimensional histogram created based onthe characteristic information from the detection unit 42, and theparticles are classified as red blood cells and platelets in the RBC/PLTmeasurement. The analysis results thus obtained are output to thedisplay unit 302 of the data processing unit 3 (step S20).

When input specifying the measurement mode is received as describedabove in step S5, the microcomputer 6 sets the parameters (operatingconditions) for the body fluid measurement, for example, the reactionchamber to use and the set time of the measurement and the like (stepS8). In the present embodiment, the measurement time is three times thetime for blood measurement, as will be described later.

The measuring unit 2 starts the pre sequence (step S10) when themeasurement mode has been switched from the previous measurement mode(in this instance, the blood measurement mode) to the body fluidmeasurement mode (step S9). The pre sequence is a process of preparingfor the body fluid measurement. Since samples which have a lowconcentration of blood cell component are measured in the body fluidmeasurement, the setting is switched from the blood measurement mode([1:NORMAL] is displayed in FIG. 8) to the body fluid measurement mode,and the lack of background influence is confirmed in the body fluidmeasurement results.

The pre sequence includes a blank check operation. The blank checkdetermination standard of the pre sequence is set at a fraction and ismore strict than the determination standard of the blank check (forexample, the blank check performed after power on and automatic wash)performed in the blood measurement mode. When the setting is changedfrom the body fluid measurement mode to the blood measurement mode, thispre sequence is not performed since there is no background influence(carry over effect) on the normal blood measurement results.Furthermore, when body fluid samples are measured in a repeated bodyfluid measurement mode, this pre sequence is not performed since thereis normally no background influence. There is concern, however, that thenext sample measurement may be affected when the body fluid sampleanalysis results exceed a predetermined value due to an extremely highnumber of particles in the body fluid since the measurement results arehigh, and therefore the operator is alerted of this concern that theanalysis results of the next sample may be affected. Then, the blankcheck measurement is performed. A configuration is desirable in which amessage “please press VERIFY” is output to the screen, and the blankcheck is performed when the operator presses the VERIFY button. In thiscase, a configuration is possible in which a CANCEL button may beprovided on the screen to transition to the standby screen withoutperforming a blank check when the operator presses the CANCEL button. Itis also desirable that a flag indicate the low reliability of themeasurement results when a blank check is not performed. Wasted reagentand time can thus be avoided by performing an additional blank checkonly when needed.

FIG. 9 is a flow chart showing the sequence of the pre sequence processperformed when the measurement mode is changed from the bloodmeasurement mode to the body fluid measurement mode. The sample analyzer1 performs the pre sequence by measuring a blank sample using themeasuring unit 2 (step S31), comparing the measurement result withpredetermined tolerance values and determining whether or not themeasurement results are less than the tolerance values using themicrocomputer 6 (step S32). When the measurement results are less thanthe tolerance values, the microcomputer 6 ends the pre sequence and theprocess returns. When the measurement results are not less than thetolerance value, the microcomputer 6 determines whether or not the blankcheck was executed the set number of times (for example, three times)(step S33), and when the number of executions of the blank check is lessthan a predetermined number, the process returns to step S31 and theblank check is performed again for the predetermined number of times.When the measurement results of the blank check performed apredetermined number of times are not less than the tolerance values, ascreen is displayed with includes a VERIFY button, BLANK CHECK button,and AUTOMATIC WASH button and the blank check measurement results aredisplayed on the display and operation unit 7 (step S34). When theoperator has pressed the VERIFY button (step S35), the microcomputer 6ends the pre sequence and the process returns. When the BLANK CHECKbutton has been pressed (step S36), the process returns to step S31 andthe blank check is performed again; when the AUOMATIC WASH button hasbeen pressed (step S37), automatic washing is performed using a specialwashing solution (strep S38), and thereafter the process returns to stepS31 and the blank check is performed again.

When the pre sequence ends as described above, the sample analyzer 1enters the standby state (step S11). When the operator presses the startswitch and starts the body fluid measurement, the sample aspirationnozzle 18 of the measuring unit 2 is immersed in the sample container inthe same manner as for the manual measurement of the blood sample. Whenthe instruction to start measurement is received by the microcomputer 6(step S12), the body fluid aspiration begins (step S13).

After the body fluid sample has been aspirated, the body fluid sample isintroduced to the sampling valve 91 in the same manner as the bloodsample. Then, the RBC/PLT measurement sample is prepared by the reactionchamber 13 (step S15). Subsequently, the DIFF measurement sample ismeasured by the white blood cell detection unit 41, and the RBC/PLTmeasurement sample is measured by the RBC/PLT detection unit 42 (stepS17). Since only the DIFF measurement sample is measured by the whiteblood cell detection unit 41 in the body fluid measurement mode, themeasurement is completed in a shorter time than the blood measurementeven though the measurement time is longer than the measurement time inthe blood measurement mode. the analysis accuracy of the low particleconcentration body fluid sample can therefore be improved by increasingthe measurement time of the body fluid measurement to be longer than themeasurement time of the blood measurement. Although the measurementaccuracy can be improved due to the increased number of particlescounted by lengthening the measurement time, a two to six fold increasein the measurement time is suitable because the sample processingability is reduced when the measurement time is excessively long, andthere is a limit to the performance of the syringe pump which deliversthe measurement sample to the white blood cell detection unit 41. In thepresent embodiment, the measurement time in the body fluid measurementmode is set at three times the measurement time of the blood measurementmode.

The RBC/PLT measurement sample is introduced to the electricalresistance detection unit 41 in the same manner for all measurementmodes, and measurement is performed under a fixed flow speed condition.The analysis processing is performed thereafter based on thecharacteristic information obtained by the measurements (step S19), andthe analysis results are output to the display unit 302 of the dataprocessing unit 3 (step S21). In the analysis processing of the bloodmeasurement mode, the DIFF scattergram and the like are analyzed, andinformation is calculated for five types of white blood cell subclasses(NEUT: neutrophil, LYMPH: lymphocyte, MONO: monocyte, EO: eosinophil,and BASO: basophil), whereas in the analysis processing of the bodyfluid measurement mode, two subclasses (MN: mononuclear cell, PMN:polymorphonuclear cell) are classified in a partially integrated formbecause there are a lesser number of blood cells and these cells aresometimes damaged. The lymphocytes and monocytes belong to mononuclearcells, and neutrophils, eosinophils, and basophils belong topolymorphonuclear cells. Since the classification algorithm is the sameas the algorithm described for the analysis processing in the bloodmeasurement mode, further description is omitted.

Next, the analysis results obtained in step S19 are compared to thetolerance value (predetermined threshold value) (step S22). Thetolerance value is the same value as the tolerance value used in theblank check of the pre sequence performed in step S10. When the analysisresult is greater than the tolerance value (step S22: Y), theverification screen 151 at the start of the blank check is displayed, asshown in FIG. 17. A message is displayed on the verification screen 151indicating there is concern that the measurement of the next sample maybe influenced due to the high measurement result. Then, the blank checkmeasurement is performed. A message display area 152 for displaying themessage “please press the VERIFY button”, a VERIFY button 153, and aCANCEL button 154 are displayed. Next, determinations are made as towhether or not the user has pressed the VERIFY button 153 or the CANCELbutton 154 (step S24), and the blank check is executed when the VERIFYbutton has been pressed (VERIFY in step S24) (step S25). The processreturns to step S5 without performing the blank check when the analysisresult obtained in step S19 is less than the tolerance value (step S22:N), and the when the CANCEL button has been pressed (CANCEL in stepS24).

Anomalous particles (macrophages, mesothelial cells, tumor cells and thelike) other than blood cells may be present in the body fluid sample.Although it is rare for such anomalous cells to be present incerebrospinal fluid, such cells appear comparatively frequently inabdominal and thoracic fluids. The influence of these anomalousparticles must be eliminated in order to obtain a high precisionclassification of blood cells within the body fluid regardless of thetype of body fluid. White blood cells in body fluid can be measured withgreater precision based on the new knowledge than anomalous particlesappear in the top part of the DIFF scattergram produced by this bloodcell analyzer of the present invention. This aspect was not consideredin the previously mentioned conventional art.

FIG. 10 is a schematic view of a scattergram obtained by measuring andanalyzing a DIFF measurement sample prepared from body fluid and whiteblood cell measurement reagent in the body fluid measurement mode of theblood cell analyzer 1 of the present embodiment. The vertical axis ofthe scattergram represents the side fluorescent light intensity (thefluorescent light intensity at the top is greatest), and the horizontalaxis represents the side scattered light intensity (the scattered lightintensity at the right side is greatest). A red blood cell ghost Gccaused by hemolysis is distributed in the region LF in which thefluorescent light intensity is weakest in the scattergram, anomalousparticles such as mesothelial cells and the like is distributed in theregion HF in which the fluorescent light intensity is greatest, andmononuclear white blood cells Mc and polynuclear white blood cells Pcare distributed in the intermediate region MF. In the analysis of thescattergram, the particle component distributed in the region MF isanalyzed as white blood cells after excluding region LF and region HF,and the particles are classified and counted in two groups. Lymphocytesand monocytes are included in the mononuclear white blood cells Mc, andneutrophils, basophils, and eosinophils are included in the polynuclearwhite blood cells Pc.

Since fewer and damaged blood cells are contained in body fluid, whiteblood cells are classified and counted as mononuclear white blood cellsand polynuclear white blood cells when analyzing white blood cells inbody fluid.

Anomalous particles (nucleated cells such as tumor cells, macrophages,mesothelial cells) other than blood cells may also be present in bodyfluid. Although it is rare for such anomalous cells to be present incerebrospinal fluid, such cells appear comparatively frequently inabdominal and thoracic fluids. In the scattergram of FIG. 10, suchnucleated cells other than white blood cells are distributed in regionHF. In the present embodiment, it is possible to determine accuratewhite blood cells counts even in body fluid which contains suchnucleated cells other than white blood cells since nucleated cells otherthan white blood cells can be identified. The degree of occurrence ofanomalous cells can be determined by counting the cells which appear inregion HF. In the present embodiment, cells are demarcated in theregions LF, MF, and HF by threshold values for demarcating each region;these threshold values may also be changed manually.

FIG. 11 compares the analysis results of the blood cell analyzer 1 ofthe present embodiment and the count results of a reference method toshow the validity of the scattergram analysis method described above.The sample material is thoracic fluid; in the drawing, “this method”refers to the white blood cell count (WBC) and anomalous particle count(Others) calculated by the blood cell analyzer 1 of the presentembodiment, and “Ref” refers to the calculation result by the referencemethods (Fuchs Rosenthal calculation method and site-spin method).Examples 1, 2, and 3 are the results of analysis of thoracic fluid inwhich anomalous particles were plentiful, and the correlation betweenthe reference methods and the analysis results of the blood cellanalyzer 1 of the present invention can be readily understood.

FIG. 13 shows a screen 200 which is displayed on the display unit 302 ofthe data processing unit 3, showing the analysis results of the DIFFmeasurement sample prepared from blood. A sample number display regionwhich displays a sample number 101 is provided at the top of the screen200, and an attribute display region which displays patient attributesis provided adjacently. The attribute display region specificallyincludes a patient ID, patient name, date of birth, sex, hospitaldepartment/ward, attending physician, date of measurement, time ofmeasurement, comments and the like. A measurement result display regionwhich displays the measurement results is provided at the bottom of theattribute display region. The measurement result display region includesseveral pages, and these pages can be displayed by selecting a pluralityof tabs 102. Tabs have a plurality of arrangements matching the mainscreen, graph screen, and measurement items. FIG. 12 is a screen whichis displayed when the graph screen tab has been selected. A graphdisplay region 104 for displaying graphs and a measurement value displayregion 103 for displaying the measurement result values are provided inthe left half of the measurement value display region, and adistribution map display region for displaying the measurement resultdistribution map 105 is provided in the right half. WBC, RBC, . . . ,NEUT#, . . . , BASO#, . . . , NEUT#, . . . , BASO % and the like, data,and units are displayed in the measurement value display region, andflagging results representing sample anomalies and disease suspicionswhich are clinically useful information relating to WBC, PLT, RBC or RETare displayed in the flag display region 104.

Six distribution maps are displayed in the distribution map displayregion 105. The scattergram on the upper left side is a DIFFscattergram. The WBC/BASO scattergram is shown at the top right, theimmature cell (IMI) scattergram is shown at mid left, and the RETscattergram is shown at mid right. The RBC scattergram is shown at thebottom left, and the PLT scattergram is shown at the bottom right.

FIG. 14 shows a screen 110 displayed in the display area 302 of the dataprocessing unit 3 as the measurement results of the DIFF measurementsample prepared from body fluid. A sample number display region 111 fordisplaying a sample number is provided at the top of the screen 110, anda patient attribute display region is provided adjacently. An [F], whichindicates measurement has been conducted in the body fluid measurementmode, is displayed at the left end of the sample number display region111. Thus, it can be clearly recognized that the analysis results arefor body fluid measurement results. The measurement result displayregion includes a plurality of pages which are selectable by tab 112. Inthis example, the tab for body fluid measurement is selected.

The measurement value display region 113 includes the name of themeasurement items for body fluid measurement rather than the measurementresults of the blood measurement mode; WBC-BF (WBC count), RBC-BF (RBCcount), MN# (mononuclear cell count (lymphocytes+monocytes)), PMN#(polymorphonuclear cell count (neutrophils+basophils+eosinophils)), MN %(ratio of mononuclear cells among white blood cells), PMN % (ratio ofpolymorphonuclear cells among white blood cells), measurement values,and units are associated and displayed. A flag display region 114 isprovided in the body fluid measurement similar to the blood measurement.Two distribution maps 115 are displayed in the distribution map displayregion, and the top scattergram is a DIFF scattergram. The bottomscattergram is an RBC scattergram.

FIG. 15 shows an example in which the Research BF tab 112 is selected inthe screen 110 of FIG. 14. This screen displays the same items as screen110 with the exception that a research parameter display region 116 isalso displayed. The research parameter display region 116 displaysnumber of particles in region HF [HF-BF#], the ratio of the number ofparticles in the region HF relative to the number of particles in theregion including both region HF and region MF [HF-BF %], and the numberof particles in the region including both region HF and region MF[TC-BF#] in FIG. 10. [HF-BF %] is the percentage of HF-BF relative toTC-BF.

FIG. 16 shows a screen 120 showing a list of stored samples which isdisplayed on the display unit 302 of the data processing unit 3.Reference number 130 refers to a patient attribute display region.Provided above this region is a measurement result display region whichdisplays the measurement result selected by a tab. A row 131 on the leftend of the measurement result display region is used to indicate whetherthe validation operation has been performed or not for the measurementresult. A “V” symbol indicates validation has been performed. A row 132on the right indicates the measurement mode. An “F” symbol indicates themeasurement results are for the body fluid mode. Although there are highvalue samples that require blank checking in the body fluid mode, andinverted “F” symbol can be displayed to indicate the blank check has notbeen performed (that is, CANCEL was selected in step S24).

Although the structure and functions of the blood cell analyzer of thepresent invention have been described as being pre-established in theblood cell analyzer, the same functions may be realized by a computerprogram so that the functions of the present invention can be realizedin a conventional blood cell analyzer by installing the computer programin a conventional blood cell analyzer.

Although the amount of sample, type of reagent, and amount of reagentare the same when preparing measurement samples for the white blood cellclassification measurement in the blood measurement mode and the whiteblood cell classification measurement in the body fluid measurement modein the present embodiment, the present invention is not limited to thisconfiguration inasmuch as the amount of sample and the amount of reagentused to prepare a measurement sample for white blood cell classificationin the body fluid measurement mode may be greater than the amount ofsample and the amount of reagent used to prepare a measurement samplefor white blood cell classification in the blood measurement mode. Sincethe measurement time is greater and the amount of measurement sampleneeded for measurement is greater for white blood cell classification inthe body fluid measurement mode than in the blood measurement mode, itis thereby possible to prepare suitable amounts of measurement samplefor white blood cell classification in the blood measurement mode andfor white blood cell classification in the body fluid measurement mode.Moreover, the type of reagent used for white blood cell classificationin the blood measurement mode may differ from the type of reagent usedfor white blood cell classification in the body fluid measurement mode.

Although white blood cell classification is performed in the body fluidmeasurement mode using scattered light and fluorescent light in thepresent embodiment, the present invention is not limited to thisconfiguration inasmuch as white blood cell classification may also beperformed in the body fluid measurement mode using, for example,scattered light and absorbed light. The measurement of absorbed lightmay be accomplished by preparing a measurement sample by mixing astaining reagent to stain the white blood cells, and other reagenttogether with the sample, supplying this measurement sample to a flowcell to form a sample flow within the flow cell, irradiating this sampleflow with light, and receiving the light emitted from the sample flowvia a photoreceptor element such as a photodiode or the like. The lightis absorbed by the white blood cells when the white blood cells passthrough the flow cell, and the degree of that absorption can be graspedas the amount of light received by the photoreceptor element. Suchmeasurement of absorbed light is disclosed in U.S. Pat. Nos. 5,122,453,and 5,138,181. furthermore, electrical resistance may be measured ratherthan scattered light, in which case white blood cells can be classifiedby the electrical resistance and absorbed light.

What is claimed is:
 1. A method for counting cells in a samplecontaining a body fluid other than blood, the method comprising: at anautomated cell counter comprising a flow cytometer, displaying a screenproviding options of measurement modes for measuring the sampleincluding a body fluid measurement mode; receiving a designation of thebody fluid measurement mode from among the measurement modes at thescreen; aspirating the sample containing the body fluid; preparing ameasurement sample by mixing the sample containing the body fluid and areagent; measuring at least light scatter and fluorescence for particlescontained in at least a part of the measurement sample by the flowcytometer; generating a scattergram from the light scatter andfluorescence, wherein generating a scattergram comprises, on the basisof the measured light scatter and fluorescence, discernibly displaying acluster of polymorphonuclear cells and a cluster of mononuclear cells inthe scattergram; and on the basis of the measured light scatter andfluorescence, counting at least one of the polymorphonuclear cells orthe mononuclear cells.
 2. The method of claim 1, wherein thepolymorphonuclear cells and mononuclear cells are discriminated frommacrophages, mesothelial cells, and tumor cells.
 3. The method of claim1, further comprising identifying nucleated cells in the scattergram. 4.The method of claim 1, further comprising calculating the proportion ofpolymorphonuclear cells in white blood cells in the body fluid and theproportion of mononuclear cells in the white blood cells.
 5. The methodof claim 1, wherein the flow cytometer is configured to form a flow ofthe measurement sample in a flow cell and irradiate the flow with lightto produce the light scatter and the fluorescence from each of theparticles in the measurement sample.
 6. The method of claim 5, whereineach of the particles in the measurement sample are plotted on thescattergram at a position addressed by the light scatter andfluorescence, the scattergram defining regions LF, MF and HF that,respectively, cover low, middle and high ranges with respect to thefluorescence, the particles in the MF is identified as white blood cellsin the body fluid, and the particles in the HF is identified asanomalous particles.
 7. The method of claim 6, wherein the particles inthe region MF is classified into the polymorphonuclear cells and themononuclear cells according to the light scatter.
 8. The method of claim6, further comprising calculating a total number of nucleated cells inthe body fluid, as sum of the number of cells in the region MF and thenumber of cells in the region HF.